Guo Shan, Yu Meng-Meng, Li Ge, Zhou Hua, Fang Feng, Shu Sai-Nan
Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Er Ke Za Zhi. 2013 Apr;51(4):260-4.
To investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.
Fresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared.
Totally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05).
Genotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.
研究人巨细胞病毒(HCMV)糖蛋白H(gH)在婴幼儿临床分离株中的基因多态性,分析gH在不同HCMV感染疾病中的基因型分布,并探讨疾病与基因型之间的相关性。
收集住院的血HCMV-IgM和HCMV-IgG阳性的不同疾病患儿的新鲜尿液标本,从中分离病毒。采用巢式聚合酶链反应(nested-PCR)结合限制性片段长度多态性(RFLP)技术对收获的临床分离株的糖蛋白H进行基因分型,纯化的PCR产物用限制性内切酶HhaI消化,消化产物通过聚丙烯酰胺凝胶电泳和银染进行基因分型,比较测序的分类和结果。
共获得102株HCMV临床分离株。这些临床分离株(43例为婴儿肝炎综合征,38例为无黄疸型肝炎,13例为肺炎,7例为血小板减少性紫癜,1例为先天性CMV感染)的糖蛋白H基因经巢式PCR检测均为阳性,阳性率为100%。结果显示,62株为gH1基因型(60.8%),40株为gH2基因型(39.2%),未观察到混合型或新基因型。在婴儿肝炎综合征(26株临床分离株为gH1基因型,17株临床分离株为gH2基因型)、无黄疸型肝炎(25株为gH1,13株为gH2)和肺炎(9株为gH1,4株为gH2)中,婴幼儿临床分离株的HCMV gH基因型分布与总体趋势一致(χ(2)=0.357,P>0.05)。然而,在血小板减少性紫癜患者的临床分离株中,gH2比gH1更常见(6株为gH2,1株为gH1,χ(2)=6.083,P<0.05)。
1型基因型是我院婴幼儿HCMV临床分离株中糖蛋白H的优势基因型。婴儿肝炎综合征、无黄疸型肝炎和肺炎中gH基因型分布无显著差异。然而,gH2是血小板减少性紫癜的优势基因型。这些发现提示gH基因型与不同临床表现之间可能存在一定相关性。